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Localization of Alu Sequences in Nuclei of Fibroblasts and Lymphocytes

A - Karyotype from a female human lymphocyte (46, XX). Chromosomes were hybridized with a probe for Alu sequences (green) and counterstained with TOPRO-3 (red). Alu sequences were used as a marker for chromosomes and chromosome bands rich in genes.
B and C - Confocal serial sections were obtained from a human G0 fibroblast nucleus (B) and a G0 lymphocyte nucleus from peripheral blood (C) after 3D FISH with the Alu probe (green) and TOPRO-3 counterstaining (red). As examples, sections made at the top, middle, and bottom of the nuclei (separated by about 1 μm) are shown from left to right. Scale bars, 5 μm.
D - Enlarged confocal mid-section through the human G0 fibroblast nucleus. Scale bar, 5 μm.
E - Enlargement of the boxed sector in (D). The color image in the middle reflects the merged images left (TOPRO-3 counterstaining, red) and right (Alu staining, green). Arrows indicate chromatin rich in Alu sequences expanding into the TOPRO-3-stained, Alu-poor nuclear rim. Scale bar, 2 μm.

Reference

<pubmed>15839726</pubmed>| PLoS Biol.

Copyright

© 2005 Bolzer et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Figure 7. doi:10.1371/journal.pbio.0030157.g007 PMID--15839726journal.pbio.0030157.g007.jpg

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current	12:25, 16 October 2014		1,095 × 1,200 (197 KB)	Z8600021 (talk \| contribs)	Figure 7. Localization of Alu Sequences in Nuclei of Fibroblasts and Lymphocytes (A) Karyotype from a female human lymphocyte (46, XX). Chromosomes were hybridized with a probe for Alu sequences (green) and counterstained with TOPRO-3 (red). Alu sequen...

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Copyright holder	Creative Commons Attribution License
Copyright status	Copyright status not set
Software used	Adobe Photoshop CS Macintosh
Short title	Figure 7
IIM version	2

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+==Localization of Alu Sequences in Nuclei of Fibroblasts and Lymphocytes==
-Figure 7. Localization of Alu Sequences in Nuclei of Fibroblasts and Lymphocytes
+* '''A''' - Karyotype from a female human lymphocyte (46, XX). Chromosomes were hybridized with a probe for Alu sequences (green) and counterstained with TOPRO-3 (red). Alu sequences were used as a marker for chromosomes and chromosome bands rich in genes.
-(A) Karyotype from a female human lymphocyte (46, XX). Chromosomes were hybridized with a probe for Alu sequences (green) and counterstained with TOPRO-3 (red). Alu sequences were used as a marker for chromosomes and chromosome bands rich in genes.
+* '''B and C''' - Confocal serial sections were obtained from a human G0 fibroblast nucleus (B) and a G0 lymphocyte nucleus from peripheral blood (C) after 3D FISH with the Alu probe (green) and TOPRO-3 counterstaining (red). As examples, sections made at the top, middle, and bottom of the nuclei (separated by about 1 μm) are shown from left to right. Scale bars, 5 μm.
-(B and C) Confocal serial sections were obtained from a human G0 fibroblast nucleus (B) and a G0 lymphocyte nucleus from peripheral blood (C) after 3D FISH with the Alu probe (green) and TOPRO-3 counterstaining (red). As examples, sections made at the top, middle, and bottom of the nuclei (separated by about 1 μm) are shown from left to right. Scale bars, 5 μm.
+* '''D''' - Enlarged confocal mid-section through the human G0 fibroblast nucleus. Scale bar, 5 μm.
-(D) Enlarged confocal mid-section through the human G0 fibroblast nucleus. Scale bar, 5 μm.
+* '''E''' - Enlargement of the boxed sector in (D). The color image in the middle reflects the merged images left (TOPRO-3 counterstaining, red) and right (Alu staining, green). Arrows indicate chromatin rich in Alu sequences expanding into the TOPRO-3-stained, Alu-poor nuclear rim. Scale bar, 2 μm.
-(E) Enlargement of the boxed sector in (D). The color image in the middle reflects the merged images left (TOPRO-3 counterstaining, red) and right (Alu staining, green). Arrows indicate chromatin rich in Alu sequences expanding into the TOPRO-3-stained, Alu-poor nuclear rim. Scale bar, 2 μm.
-doi:10.1371/journal.pbio.0030157.g007
+{{cell nuclear DNA links}}
+===Reference===
+<pubmed>15839726</pubmed>| [http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.0030157 PLoS Biol.]
+====Copyright====
-PLoS Biol. 2005 May;3(5):e157. Epub 2005 Apr 26.
+© 2005 Bolzer et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
-Three-dimensional maps of all chromosomes in human male fibroblast nuclei and prometaphase rosettes.
-Bolzer A1, Kreth G, Solovei I, Koehler D, Saracoglu K, Fauth C, Müller S, Eils R, Cremer C, Speicher MR, Cremer T.
-Author information
-Abstract
-Studies of higher-order chromatin arrangements are an essential part of ongoing attempts to explore changes in epigenome structure and their functional implications during development and cell differentiation. However, the extent and cell-type-specificity of three-dimensional (3D) chromosome arrangements has remained controversial. In order to overcome technical limitations of previous studies, we have developed tools that allow the quantitative 3D positional mapping of all chromosomes simultaneously. We present unequivocal evidence for a probabilistic 3D order of prometaphase chromosomes, as well as of chromosome territories (CTs) in nuclei of quiescent (G0) and cycling (early S-phase) human diploid fibroblasts (46, XY). Radial distance measurements showed a probabilistic, highly nonrandom correlation with chromosome size: small chromosomes-independently of their gene density-were distributed significantly closer to the center of the nucleus or prometaphase rosette, while large chromosomes were located closer to the nuclear or rosette rim. This arrangement was independently confirmed in both human fibroblast and amniotic fluid cell nuclei. Notably, these cell types exhibit flat-ellipsoidal cell nuclei, in contrast to the spherical nuclei of lymphocytes and several other human cell types, for which we and others previously demonstrated gene-density-correlated radial 3D CT arrangements. Modeling of 3D CT arrangements suggests that cell-type-specific differences in radial CT arrangements are not solely due to geometrical constraints that result from nuclear shape differences. We also found gene-density-correlated arrangements of higher-order chromatin shared by all human cell types studied so far. Chromatin domains, which are gene-poor, form a layer beneath the nuclear envelope, while gene-dense chromatin is enriched in the nuclear interior. We discuss the possible functional implications of this finding.
-PMID 15839726
+Figure 7. doi:10.1371/journal.pbio.0030157.g007 PMID--15839726journal.pbio.0030157.g007.jpg
-http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.0030157
+[[Category:Human]][[Category:Adult]][[Category:Female]]
-PMID--15839726journal.pbio.0030157.g007.jpg